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Day 1: TWIN-SHIME inoculation with the same faecal sample.
Days 1- 22: Stabilitation period.
Days 1t- 11t: Treatment period. Supplementation of Provinols ® and Provinols ® + L. plantarum IFPL935
Day 12: End of supplementation.
Days 12-20: Washout period.
E. Barroso1*, T. van de Wiele2, A. Jiménez-Girón1, I. Muñoz-González1, M.V. Moreno-Arribas1, B. Bartolomé1, C. Peláez1, M.C Martínez-Cuesta1, T. Requena1
Lactobacillus plantarum IFPL935 improves colonic metabolism in a simulator of the human intestinal microbial ecosystem
during feeding with red wine phenolic compounds
1Instituto de Investigación en Ciencias de la Alimentación CIAL (CSIC-UAM), Madrid, Spain.2LabMET, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.*[email protected]
In the present work we have evaluated the adaptation of the colonic microbiota developed in two parallel five-stage in vitro gastrointestinal simulators (TWIN-SHIME) to long-term feeding with a polyphenol-rich red wine extract and the combination of this extract with a potential probiotic strain Lactobacillus plantarum IFPL935.
In the present work we have evaluated the adaptation of the colonic microbiota developed in two parallel five-stage in vitro gastrointestinal simulators (TWIN-SHIME) to long-term feeding with a polyphenol-rich red wine extract and the combination of this extract with a potential probiotic strain Lactobacillus plantarum IFPL935.
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20 22 1t 2t 3t 4t 5t 9t 11t 12 13 16 19
Provinols + L. plantarum IFPL 935
AC Provinols
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20 22 1t 2t 3t 4t 5t 9t 11t 12 13 16 19
TC Provinols + L. plantarum IFPL 935
TC Provinols
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20 22 1t 2t 3t 4t 5t 9t 11t 12 13 16 19
DC Provinols + L. plantarum IFPL 935
DC Provinols
DaysDays Days
Bu
tyra
te (
mg
/mL
)
AC TC DC
HO
HO
O
OH
OH
OHHO
HO OH
OH
OHOH
(+) – catechin
4-hydroxy-5-(3’,4’-dihydroxyphenyl)-valeric acid
1-(3’,4’-dihydroxyphenyl)-3-(2’’,4’’,6’’-trihydroxyphenyl)-propan-2-ol
OH
OHHOOC OH
4-Hydroxy-5-(phenyl)-valeric acid
L. plantarum IFPL935(Tabasco et al., 2011)
Co
ntr
ol:
pH
, Tª,
fee
d
Experimental period
• The polyphenolic extract addition (day 1t) caused a sharp reduction of the fermentation activity (SCFAs) in the three colon vessels.
• Significant increase of butyrate production during the polyphenols treatment and the washout period in vessels TC and DC treated with L. plantarum IFPL935 compared with the vessels without the strain.
BUTYRATE PRODUCTION
This work was supported by Research Projects AGL2009-13361-C02-00, RM2011-00003-00-00 and Consolider Ingenio 2010 FUN-C-FOOD-CSD2007-00063, and program ALIBIRD P2009/AGR-1469.
MICROBIAL CATABOLISM OF WINE POLYPHENOLS
Catabolism of (+)-catechin.
• The evolution of the content of (+)-catechin during the treatment and washout periods shows disappearance of this compound in TC and DC, in accordance to the progressive appearance of phenolic metabolites starting after 4-5 days of treatment.
• L. plantarum tend to favor the catabolism of (+)-catechin, showing a more rapid disappearance of the precursor to the formation of further-intermediate products.
• A decrease in the microbial community is observed the first days of treatment, related with the reduction of butyrate and total SCFAs (data not shown).
• The increase of butyrate was associated with a faster recovery of the butyrate-producing bacteria, Clostridium leptum, Clostridium coccoides-Eubacterium rectale group and Ruminococcus on TC and DC vessels in the SHIME supplemented with L. plantarum IFPL935.
Conclusion L. plantarum IFPL935 shows a protective effect on butyrate-producing bacteria and is able to improve colonic metabolism by promoting the formation of biologically significant compounds
Conclusion L. plantarum IFPL935 shows a protective effect on butyrate-producing bacteria and is able to improve colonic metabolism by promoting the formation of biologically significant compounds
AC
TC
DC
0
0,2
0,4
0,6
0,8
0 2 4 6 8 10 12 14 16 18 20
Days
0
0,2
0,4
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0,8
0 2 4 6 8 10 12 14 16 18 20
Days
0
0,2
0,4
0,6
0,8
0 2 4 6 8 10 12 14 16 18 20
Days
00,020,040,060,08
0,1
0 2 4 6 8 10 12 14 16 18 20
Days
0
0,02
0,04
0,06
0,08
0,1
0 2 4 6 8 10 12 14 16 18 20
Days
4-H
ydro
xy-5
-(3’,4
’-dih
ydro
xyph
enyl
)-val
eric
acid
(µg/
mL)
0
0,02
0,04
0,06
0,08
0,1
0 2 4 6 8 10 12 14 16 18 20
Days
(+)-
Cate
chin
(µg/
mL)
0
20
40
60
80
0 2 4 6 8 10 12 14 16 18 20
Days
4-H
ydro
xy-5
-(ph
enyl
)-val
eric
acid
(µg/
mL)
020406080
0 2 4 6 8 10 12 14 16 18 20
Days
0
20
40
60
80
0 2 4 6 8 10 12 14 16 18 20
Days
Treatment Washout Treatment Washout Treatment Washout
MICROBIAL COUNTS by qPCR(counts are expressed in average log copy number/mL)
Provinols®Provinols® + L. plantarum IFPL935
Provinols®Provinols® + L. plantarum IFPL935
Bacteria l group Compartment Time period
AC Stab. 7,32 (0,21) 7,06 (0,14)Treat. days 1-5 7,06 (0,40) 7,34 (0,26)Treat. days 9-11 6,86 (0,20) 7,00 (0,22)Washout 7,17 (0,21) 7,20 (0,12)
TC Stab. 7,60 (0,09) 7,42 (0,21)Treat. days 1-5 7,26 (0,25) 6,93 (0,48)Treat. days 9-11 7,06 (0,36) 7,19 (0,08)Washout 7,48 (0,01) 7,51 (0,27)
DC Stab. 7,43 (0,09) 7,34 (0,18)Treat. days 1-5 7,11 (0,38) 6,86 (0,59)Treat. days 9-11 7,34 (0,35) 7,33 (0,04)Washout 7,57 (0,16) 7,24 (0,31)
AC Stab. 6,79 (0,29) 6,57 (0,44)Treat. days 1-5 5,94 (0,31) 6,23 (0,35)Treat. days 9-11 6,12 (0,47) 6,16 (0,40)Washout 5,91 (0,45) 6,73 (0,10)
TC Stab. 6,63 (0,08) 6,51 (0,09)Treat. days 1-5 5,84 (0,18) 6,01 (0,51)Treat. days 9-11 6,29 (0,18) 6,74 (0,02)Washout 6,37 (0,10) 6,68 (0,07)
DC Stab. 6,31 (0,01) 6,44 (0,08)Treat. days 1-5 5,68 (0,23) 5,58 (0,50)Treat. days 9-11 6,21 (0,29) 6,55 (0,02)Washout 6,49 (0,14) 6,45 (0,35)
AC Stab. 5,74 (0,28) 5,73 (0,42)Treat. days 1-5 4,94 (0,42) 4,19 (0,51)Treat. days 9-11 5,94 (0,15) 5,74 (0,00)Washout 6,19 (0,09) 5,59 (0,16)
TC Stab. 5,97 (0,12) 5,72 (0,10)Treat. days 1-5 5,66 (0,20) 4,93 (0,53)Treat. days 9-11 6,31 (0,41) 6,23 (0,18)Washout 6,59 (0,21) 6,19 (0,20)
DC Stab. 6,03 (0,32) 5,69 (0,21)Treat. days 1-5 5,48 (0,13) 4,96 (0,49)Treat. days 9-11 6,23 (0,30) 6,13 (0,32)Washout 6,33 (0,02) 6,18 (0,45)
AC Stab. 6,31 (0,14) 6,20 (0,05)Treat. days 1-5 5,37 (0,42) 5,09 (0,38)Treat. days 9-11 6,27 (0,10) 6,06 (0,04)Washout 6,18 (0,13) 6,02 (0,15)
TC Stab. 6,54 (0,07) 6,26 (0,14)Treat. days 1-5 6,04 (0,06) 5,98 (0,17)Treat. days 9-11 6,14 (0,45) 6,34 (0,19)Washout 6,28 (0,03) 6,28 (0,20)
DC Stab. 6,19 (0,26) 6,25 (0,10)Treat. days 1-5 5,70 (0,20) 5,62 (0,57)Treat. days 9-11 5,66 (0,60) 6,38 (0,21)Washout 6,32 (0,07) 6,10 (0,26)
AC Stab. 3,66 (0,21) 2,78 (0,26)Treat. days 1-5 2,91 (0,41) 1,90 (0,61)Treat. days 9-11 3,70 (0,31) 1,39 (0,09)Washout 3,76 (0,12) 1,80 (0,07)
TC Stab. 5,33 (0,10) 5,18 (0,05)Treat. days 1-5 4,35 (0,27) 4,33 (0,33)Treat. days 9-11 4,60 (0,30) 5,18 (0,27)Washout 5,15 (0,19) 5,16 (0,04)
DC Stab. 5,40 (0,13) 5,40 (0,13)Treat. days 1-5 4,62 (0,32) 4,55 (0,64)Treat. days 9-11 4,83 (0,49) 5,43 (0,23)Washout 5,34 (0,26) 5,17 (0,21)
AC Stab. 3,23 (0,15) 2,72 (0,20)Treat. days 1-5 2,75 (0,24) 2,84 (0,28)Treat. days 9-11 3,23 (0,25) 2,25 (0,08)Washout 3,30 (0,14) 2,49 (0,13)
TC Stab. 5,15 (0,15) 5,10 (0,06)Treat. days 1-5 4,28 (0,42) 4,24 (0,46)Treat. days 9-11 4,34 (0,19) 4,65 (0,38)Washout 4,86 (0,54) 4,79 (0,16)
DC Stab. 6,00 (0,14) 5,95 (0,05)Treat. days 1-5 5,27 (0,38) 5,05 (0,70)Treat. days 9-11 5,45 (0,46) 5,47 (0,39)Washout 5,45 (0,66) 5,62 (0,11)
Tota l Bacteria
Provinols Provinols + L. plantarum IFPL935
Ruminococccus spp.
Clostridium leptum
Clostridium coccoides-Eubacterium rectale group
Bifidobacterium spp.
Lactobacillus spp.
Reference:Tabasco, R., Sánchez-Patán, F., Monagas, M., Bartolomé, B., Moreno-Arribas, M.V., Peláez, C., Requena, T. 2011 Effect of grape polyphenols on lactic acid bacteria and bifidobacteria growth:resistance and metabolism. Food Microbiol. 28: 1345-1352.
